KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
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| Antibodies | ||
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| V5 tag (D3H8Q) | Cell Signaling Technology | Cat# 13202; Lot 4 RRID: AB_2687461 |
| KMT2D K1330me1/me2 | Eurogentec | Custom |
| KMT2D pS1331 | Eurogentec | Custom |
| Vinculin | Cell Signaling Technology | Cat# 4650 RRID: AB_10559207 |
| KMT2D | Fortis LifeSciences | Cat# A300-BL1185, RB X-ALR |
| phospho-AKT (Ser473) (D9E) | Cell Signaling Technology | Cat# 4060 RRID: AB_2315049 |
| phospho-RXRXX(S/T) (23C8D2) | Cell Signaling Technology | Cat# 10001 RRID: AB_10950819 |
| HA tag (C29F4) | Cell Signaling Technology | Cat# 3724 RRID: AB_1549585 |
| SMYD2 (D14H7) | Cell Signaling Technology | Cat #9734 RRID: AB_10889559 |
| DYKDDDDK Tag (D6W5B) Rabbit mAb | Cell Signaling Technology | Cat #14793; Lot 7 RRID: AB_2572291 |
| ECL™ Anti-Rabbit IgG Horseradish Peroxidase linked whole antibody (from donkey) | GE Healthcare | Cat# NA934V; Lot 17010251 RRID: AB_772206 |
| KMT2D ChIP antibody | Sigma Aldrich | Cat# HPA035877 RRID: AB_10670673 |
| V5 antibody | Abcam | Cat#ab9116 RRID:AB_307024 |
| H3 Antibody | Abcam | Cat #ab1791 RRID: AB_302613 |
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| Bacterial and virus strains | ||
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| NEB®5-alpha Competent E. coli (High Efficiency) | NEB | Cat# C2987H |
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| Chemicals, peptides, and recombinant proteins 3H-SAM |
Perkin Elmer | NET155V250UC |
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| Ultima Gold Scintillation Cocktail | Revvity | Cat# 6013321 |
| α-Cyano-4-hydroxycinnamic acid | Sigma Aldrich | Cat#C8982 |
| KMT2D peptides (unmodified, K1330me1, K1330me2, pS1331) Sequence: AHGGRGRGRARL KSTASSIETLVVAD |
GenScript | Custom |
| V5-agarose beads | Millipore Sigma | Cat# A7345 |
| EZviewTM RED Anti-HA agarose beads | Millipore Sigma | Cat# E6779 |
| H3 peptide (aa1–21) | Genscript | Custom |
| MK2206 | Selleckchem | Catalog No.S1078 |
| Alpelisib | Medchem Express | Cat# HY-15244 |
| Capivasertib | AstraZeneca | N/A |
| Fulvestrant | Selleckchem | Cat# S1191 |
| LLY-507 | Sigma Aldrich | Cat# SML1279 |
| AZ506 | Medchem Express | Cat# HY-134828 |
| EPZ033294 | Viva Biotech | N/A |
| AZ Compound 39 | Astra Zeneca | N/A |
| Doxycycline hydrochloride Dimethyl Sulfoxide |
Millipore Sigma Corning |
Cat #D3447 Cat# 25–950-CQC; Lot 01019003 |
| Crystal Violet | Sigma Aldrich | Cat #V5265 |
| PrestoBlue | ThermoFisher Scientific | Cat #A13262 |
| Y-27632 | Tocris | Cat# 1254 |
| TrypLE | ThermoFisher Scientific | Cat# 12605036 |
| Protein G Dynabeads | ThermoFisher Scientific | Cat #10004D |
| AMPure beads XP | Beckman Coulter | A63880 |
| SYBR™ Select Master Mix for CFX | Applied Biosystems | Cat# 4472942; Lot 00815825 |
| Matrigel | Corning | Cat# 354262 |
| Lipofectamine 3000 | ThermoFisher Scientific | Cat# L3000015 |
| Recombinant AKT1 | SignalChem | Cat#A16–10G |
| Recombinant AKT2 | SignalChem | Cat #A17–10H |
| Recombinant AKT3 | SignalChem | Cat #A18–10G |
| Recombinant SGK1 | SignalChem | Cat #S06–10G |
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| Critical commercial assays | ||
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| QuikChange Lightning Site-Directed Mutagenesis Kit |
Agilent | Cat#210518 |
| Q5 mutagenesis kit | NEB | Cat #E0552S |
| Roche MTT kit | Millipore Sigma | Cat# 11465007001 |
| Cell-Titer Glo 3D | Promega | Cat# G9681 |
| Qiagen RNeasy® Mini Kit (250) | Qiagen | Cat# 74106; Lot 163029673 |
| iScript™ cDNA Synthesis Kit | Bio-Rad | Cat# 1708891 |
| CUTANA CUT & RUN Kit | EpiCypher | Cat# 14–1048 |
| CUTANA CUT & RUN Library Prep Kit | EpiCypher | Cat# 14–1001 |
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| Deposited data | ||
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| ChIP-seq/RNA-seq/Cut&Run fastq files | This paper | GSE255295, GSE253957, GSE253958 |
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| Experimental models: Cell lines | ||
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| h-MCF7 | ATCC | ATCC® HTB-22™ |
| h-T47D | ATCC | ATCC® HTB-133™ |
| h-293T | ATCC | ATCC® CRL-3216™ |
| HCI-011 | Welm Laboratory | N/A |
| HCI-017 | Welm Laboratory | N/A |
| HCI-018 | Welm Laboratory | N/A |
| Homozygous KMT2D K1330A (AAG>gcG) clones | Synthego | Custom |
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| Experimental models: Organisms/strains | ||
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| Female NSG mice | Antitumor assessment core (MSKCC) | N/A |
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| Oligonucleotides | ||
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|
h-GREB1 F: GACAGGACTGTGGACTGGTG R: ATTTGTTTCCAGCCCTCCTT |
IDT | N/A |
|
h-TFF1 F: TTGTGGTTTTCCTGGTGTCA R: CCGAGCTCTGGGACTAATCA |
IDT | N/A |
|
h-WISP2 F: CAGGGGTCGCAGTCCACAAAA R: AGGCAGTGAGTTAGAGGAAAGG |
IDT | N/A |
|
h-GAPDH
F: AACAGCGACACCCATCCTG R: CATACCAGGAAATGAGCTTGACAA SMYD2 Y240A Mutagenesis Primers F: TTTTACCAGCGCCATTGATCTC CTGTACCCAACG R: ACCTCCTCTCCCGGCTTG |
IDT | N/A |
| SMYD2 Y240A Mutagenesis Primers F: TTTTACCAGCGCCATTGATCTC CTGTACCCAACG R: ACCTCCTCTCCCGGCTTG |
IDT | N/A |
| KMT2D Y5426A Mutagenesis Primers F:GCACACAATGGTTATCGAGGCCAT TGGCACCATCATTCGG R:CCGAATGATGGTGCCAATGGCCT CGATAACCATTGTGTGC |
IDT | N/A |
| KMT2D K1330A Mutagenesis Primers F:AGGACGGGCCCGGCTAGCGTC AACTGCTTCTTC R:GAAGAAGCAGTTGACGCTAGC CGGGCCCGTCCT |
IDT | N/A |
| KMT2D K1330R Mutagenesis Primers F:GACGGGCCCGGCTAAGGTC AACTGCTTCTT R:AAGAAGCAGTTGACCTTAGCC GGGCCCGTC |
IDT | N/A |
| shSMYD2–1 Antisense Guide Sequence TTTGATCTCAGAAATATATGGA |
This paper | N/A |
| shSMYD2–2 Antisense Guide Sequence TAAATGAAAACTGAAATGCTGC |
This paper | N/A |
|
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| Recombinant DNA | ||
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| plx302 V5-tagged KMT2D | Toska et al., 20174 | N/A |
| pCMV HA-tagged KMT2D | Zhang et al., 20159 | N/A |
| pcDNA3.1 FLAG SMYD2 | Generated in the Luo Laboratory | N/A |
| LT3 REPIR | MSKCC RNAi core | N/A |
| pCMV-VSVG | Stewart et al., 200329 | Addgene 8454 |
| pCMV-dR8.2 | Stewart et al., 200329 | Addgene 8455 |
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| Software and algorithms | ||
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| Gen5 software v1.02 | Agilent | https://www.agilent.com/en/product/cell-analysis/cell-imaging-microscopy/cell-imaging-microscopy-software/biotek-gen5-software-for-imaging-microscopy-1623226 |
| Adobe Illustrator 2024 | Adobe | https://www.adobe.com/products/illustrator/free-trial-download.html |
| Graphpad Prism v9.4.1 | Graphpad Software, Inc. | https://www.graphpad.com/scientific-software/prism/ |
| cBioportal | Cerami et al., 201230 Gao et al., 201331 de Bruijn et al., 202332 |
https://www.cbioportal.org/ |
| FastQC v0.11.9 | Andrews, S. (2010) | https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ |
| Salmon v1.10.0 | Patro et al., 201733 | https://github.com/COMBINE-lab/Salmon |
| DESeq2 (v1.28.1) | Love et al., 201434 | http://www.bioconductor.org/packages/release/bioc/html/DESeq2.html |
| GSEA software v4.3.2 | Subramanian, Tamayo et al., 200535 and Mootha, Lindgren et al. 200336 |
https://www.gsea-msigdb.org/gsea/index.jsp |
| MSigDB (v7.0) | Subramanian et al., 200535; Liberzon et al., 201537 |
http://software.broadinstitute.org/gsea/msigdb |
| bowtie2 v2.4.2 bedtools v2.30.0 |
Ben Langmead & Steven Salzberg, 201238 Quinian and Hall, 201039 |
https://www.nature.com/articles/nmeth.1923) https://github.com/arq5x/bedtools2 |
| MACS2 v2.1.1 | Zhang et al., 200840 | https://liulab-dfci.github.io/software/ |
| GenomicAlignments v1.36.0 | Lawrence M et al., 201341 | https://bioconductor.org/packages/release/bioc/html/GenomicAlignments.html |
| deepTools | Ramirez et al., 201642 | http://deeptools.ie-freiburg.mpg.de/ |
| HOMER v4.11 | Heinz et al., 201043 | |
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| Other | ||
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| Tri-Carb 2910TR liquid scintillation counter | Perkin Elmer | N/A |
| BioTek Cytation 5 microplate reader | Agilent | N/A |
| Omega FLUOstar luminometer | BMG Labtech | BMG-415–103 |
| QuantStudio™ 5 | Applied Biosystems | A28570 |